CN116377009A - Ginsenoside extract and preparation method and application thereof - Google Patents
Ginsenoside extract and preparation method and application thereof Download PDFInfo
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P33/00—Preparation of steroids
- C12P33/20—Preparation of steroids containing heterocyclic rings
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/63—Steroids; Derivatives thereof
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
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- A61P17/00—Drugs for dermatological disorders
- A61P17/14—Drugs for dermatological disorders for baldness or alopecia
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- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/02—Preparations for care of the skin for chemically bleaching or whitening the skin
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
- A61K2800/805—Corresponding aspects not provided for by any of codes A61K2800/81 - A61K2800/95
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- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
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Abstract
The invention belongs to the technical field of ginsenoside extraction, and provides a ginsenoside extract, a preparation method and application thereof, wherein the preparation method of the ginsenoside extract comprises the following steps: (1) Drying ginseng, pulverizing, mixing with glycosidase solution, and performing enzymolysis to obtain enzymolysis solution; (2) Sterilizing the enzymolysis liquid, mixing with a composite microbial inoculum, and fermenting to obtain the ginsenoside extract. The ginsenoside extract prepared by the preparation method contains various rare ginsenosides, can be prepared into products with effects of relieving and repairing, resisting wrinkles and whitening, preventing hair loss and growing hair, and provides a new idea for the cosmetic industry.
Description
Technical Field
The invention relates to the technical field of ginsenoside extraction, in particular to a ginsenoside extract and a preparation method and application thereof.
Background
The Chinese medicinal ginseng is root of Panax ginseng of Araliaceae, and has effects of invigorating primordial qi, invigorating spleen, nourishing lung, promoting salivation, quenching thirst, tranquilizing mind, improving intelligence, etc. The ginseng contains a plurality of active ingredients, wherein ginsenoside is one of the most main ingredients of the ginseng for playing the role. The ginsenoside has high content of Rb1, rb2, rc, rd, re and Rg1, but has low biological activity and low absorptivity in human body. The rare ginsenoside with higher biological activity has low content and is not easy to extract, and an additional preparation method is generally required. The current methods for preparing rare ginsenoside are 3: the main components in the ginsenoside are converted into rare saponins by microbial fermentation; the microorganisms such as escherichia coli, yeast and the like are modified by utilizing metabolic engineering and synthetic biology to directly produce rare ginsenoside; the main saponins are converted into rare saponins by glycosidases. However, the three preparation methods have complex processes, require a large amount of manpower and material resources, and are not beneficial to industrial production. Therefore, it is needed to provide a preparation method of rare saponins with simple process.
Disclosure of Invention
The invention aims to provide a ginsenoside extract, a preparation method and application thereof, wherein after ginseng is subjected to enzymolysis and fermentation, the extract contains various rare ginsenosides, and the ginsenoside extract can be prepared into products with effects of relieving and repairing, resisting wrinkles and whitening, preventing hair loss and generating hair.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides a preparation method of a ginsenoside extract, which comprises the following steps:
(1) Drying ginseng, pulverizing, mixing with glycosidase solution, and performing enzymolysis to obtain enzymolysis solution;
(2) Sterilizing the enzymolysis liquid, mixing with a composite microbial inoculum, and fermenting to obtain the ginsenoside extract.
Preferably, the temperature of the drying in the step (1) is 40-50 ℃, and the time of the drying is 2-6 h.
Preferably, the ratio of ginseng to glycosidase solution in step (1) is 1g: 5-10 mL;
the glycosidase solution comprises one or more of beta-glucosidase, beta-galactosidase, alpha-arabinofuranosidase and beta-xylosidase;
the concentration of the enzyme in the glycosidase solution is independently 3-10U/mL.
Preferably, the temperature of the enzymolysis in the step (1) is 40-50 ℃, and the enzymolysis time is 20-40 min.
Preferably, the pH value of the solution is adjusted to 4.5-5.5 before enzymolysis.
Preferably, the compound microbial inoculum in the step (2) comprises lactobacillus fermentum and lactobacillus rhamnosus, and the mass ratio of the lactobacillus fermentum to the lactobacillus rhamnosus is 1:1-3.
Preferably, the inoculation amount of the composite microbial inoculum in the step (2) is 5-10%, the fermentation temperature is 35-40 ℃, and the fermentation time is 24-48 h.
The invention also provides the ginsenoside extract prepared by any one of the preparation methods.
The invention also provides application of the ginsenoside extract in preparing products with effects of relieving and repairing, resisting wrinkles and whitening, preventing hair loss and promoting hair growth.
Preferably, the types of the products include facial cleanser, essence cream, face cream, facial mask, hair conditioner, shampoo, body cream.
Compared with the prior art, the invention has the beneficial effects that:
the invention enriches rare saponins in ginseng after enzymolysis and fermentation, the obtained ginsenoside extract contains various rare saponins, the preparation method is simple, and the obtained extract has high active ingredient content and can be used for preparing cosmetics with effects of relieving and repairing, resisting wrinkles and whitening, preventing hair loss and growing hair.
Detailed Description
The invention provides a preparation method of a ginsenoside extract, which comprises the following steps:
(1) Drying ginseng, pulverizing, mixing with glycosidase solution, and performing enzymolysis to obtain enzymolysis solution;
(2) Sterilizing the enzymolysis liquid, mixing with a composite microbial inoculum, and fermenting to obtain the ginsenoside extract.
In the invention, firstly, ginseng is dried and crushed, and then mixed with glycosidase solution for enzymolysis to obtain enzymolysis liquid, wherein the drying temperature is 40-50 ℃, preferably 42-48 ℃, and more preferably 44-46 ℃; the drying time is 2-6 hours, preferably 3-5 hours, and more preferably 3.5-4.5 hours; the drying method is achieved by adopting a conventional drying method in the field.
In the present invention, the pulverizing method may be a conventional pulverizing method in the art.
In the invention, the ratio of the ginseng to the glycosidase solution is 1g: 5-10 mL, preferably 1g:6 to 9mL, more preferably 1g: 7-8 mL; the glycosidase solution comprises one or more of beta-glucosidase, beta-galactosidase, alpha-arabinofuranosidase and beta-xylosidase; the concentration of each enzyme in the glycosidase solution is independently 3 to 10U/mL, preferably 4 to 8U/mL, and more preferably 5 to 6U/mL.
In the invention, the enzymolysis temperature is 40-50 ℃, preferably 42-48 ℃, and more preferably 44-46 ℃; the enzymolysis time is 20 to 40min, preferably 25 to 35min, and more preferably 28 to 32min.
In the present invention, the pH of the solution is adjusted to 4.5 to 5.5, preferably 4.7 to 5.3, more preferably 4.9 to 5.1, before the enzymatic hydrolysis is performed.
In the invention, the enzymolysis liquid is sterilized and then mixed with a composite microbial agent for fermentation to obtain a ginsenoside extract, wherein the composite microbial agent comprises lactobacillus fermentum and lactobacillus rhamnosus, and the mass ratio of the lactobacillus fermentum to the lactobacillus rhamnosus is 1:1-3, preferably 1:1.5-2.5, and further preferably 1:1.8-2.2; the viable count in the lactobacillus fermentum and the lactobacillus rhamnosus is 10 independently 8 ～10 9 CFU/g, preferably 1.2X10 8 ～1.8×10 8 CFU/g, more preferably 1.4X10 8 ～1.6×10 8 CFU/g。
In the invention, the sterilization temperature is 120-130 ℃, preferably 122-128 ℃, and more preferably 124-126 ℃; the sterilization time is 20 to 30 minutes, preferably 22 to 28 minutes, and more preferably 24 to 26 minutes.
In the present invention, the inoculation amount of the composite microbial inoculum is 5 to 10%, preferably 6 to 9%, and more preferably 7 to 8%; the fermentation temperature is 35-40 ℃, preferably 36-39 ℃, and more preferably 37-38 ℃; the fermentation time is 24 to 48 hours, preferably 30 to 45 hours, and more preferably 35 to 40 hours.
In the present invention, a centrifugation step is further included after the completion of fermentation to separate the cells, wherein the rotational speed of the centrifugation is 1000 to 2000rpm, preferably 1200 to 1800rpm, and more preferably 1400 to 1600rpm; the centrifugation time is 5 to 10 minutes, preferably 6 to 9 minutes, and more preferably 7 to 8 minutes.
In the invention, sterilization is carried out after centrifugal treatment, wherein the sterilization temperature is 120-130 ℃, preferably 122-128 ℃, and more preferably 124-126 ℃; the sterilization time is 20 to 30 minutes, preferably 22 to 28 minutes, and more preferably 24 to 26 minutes.
The invention also provides the ginsenoside extract prepared by any one of the preparation methods.
The invention also provides application of the ginsenoside extract in preparing products with effects of relieving and repairing, resisting wrinkles and whitening, preventing hair loss and promoting hair growth.
In the present invention, the types of the products include face washes, essences, essence creams, face masks, hair conditioners, shampoos, body milks.
The technical solutions provided by the present invention are described in detail below with reference to examples, but they should not be construed as limiting the scope of the present invention.
Example 1
The embodiment provides a ginsenoside extract, and the preparation method comprises the following steps:
(1) Taking cleaned ginseng, putting the cleaned ginseng into a drying box, drying at 45 ℃ for 5 hours, and crushing to obtain ginseng powder.
(2) Preparing 5U/mL glycosidase solution of beta-glucosidase and beta-galactosidase (all purchased from Shanghai Michelia Biochemical technology Co., ltd.), mixing 100g of ginseng powder with 800mL glycosidase solution, regulating pH to 5.0, performing enzymolysis at 45deg.C for 30min to obtain enzymolysis solution, and sterilizing at 121deg.C for 20 min.
(3) The viable count is 10 8 Mixing cfu/g lactobacillus fermentum and lactobacillus rhamnosus (both purchased from China general microbiological culture collection center) according to a mass ratio of 1:2, inoculating into the fermentation broth with an inoculum size of 5%, fermenting at 37 ℃ for 36 hours to obtain ginsenoside fermentation broth, centrifuging the fermentation broth at 1000rpm for 10min, and sterilizing at 121 ℃ for 20min to obtain the ginsenoside extract.
Example 2
The embodiment provides a ginsenoside extract, and the preparation method comprises the following steps:
(2) Taking the cleaned ginseng, putting the cleaned ginseng into a drying box, drying the ginseng for 6 hours at 40 ℃, and crushing the ginseng to obtain ginseng powder.
(2) Preparing glycosidase solution with independent concentration of 6U/mL from beta-glucosidase, beta-galactosidase and beta-xylosidase (all purchased from Shanghai Michelin Biochemical technology Co., ltd.), mixing 100g of ginseng powder with 500mL of glycosidase solution, regulating pH to 4.8, performing enzymolysis at 40deg.C for 40min to obtain enzymolysis solution, and sterilizing at 121deg.C for 20 min.
(3) The viable count is 10 9 Mixing cfu/g lactobacillus fermentum and lactobacillus rhamnosus (both purchased from China general microbiological culture collection center) according to a mass ratio of 1:1, inoculating 8% of the mixture into the fermentation broth, fermenting at 35 ℃ for 48 hours to obtain ginsenoside fermentation broth, centrifuging the fermentation broth at 1000rpm for 10min, and sterilizing at 121 ℃ for 20min to obtain the ginsenoside extract.
Example 3
The embodiment provides a ginsenoside extract, and the preparation method comprises the following steps:
(3) Taking cleaned ginseng, putting the cleaned ginseng into a drying box, drying the ginseng for 4 hours at 50 ℃, and crushing the ginseng to obtain ginseng powder.
(2) Preparing glycosidase solution with independent concentration of 10U/mL from beta-galactosidase, alpha-arabinopyranose and beta-xylosidase (all purchased from Shanghai Michelia Biochemical technology Co., ltd.), mixing 100g of ginseng powder with 1000mL of glycosidase solution, regulating pH to 5.2, performing enzymolysis at 45deg.C for 30min to obtain enzymolysis solution, and sterilizing at 121deg.C for 20 min.
(3) The viable count is 10 9 Mixing cfu/g lactobacillus fermentum and lactobacillus rhamnosus (both purchased from China general microbiological culture collection center) according to a mass ratio of 1:3, inoculating into the fermentation broth with an inoculum size of 6%, fermenting at 38deg.C for 36h to obtain ginsenoside fermentation broth, centrifuging the fermentation broth at 1000rpm for 10min, and sterilizing at 121deg.C for 20min to obtain ginsenoside extract.
Comparative example 1
This comparative example was different from example 1 in that the subsequent fermentation treatment was not performed after obtaining the ginseng enzymatic hydrolysate, and the remaining parameters were the same as those of example 1.
Comparative example 2
This comparative example was different from example 1 in that the ginseng powder was directly mixed with 8 times of water and then fermented without performing the enzymolysis treatment after obtaining the ginseng powder, and the other parameters were the same as those of example 1.
Comparative example 3
The comparative example is different from example 1 in that the composite microbial inoculum used for fermentation is lactobacillus fermentum and lactobacillus rhamnosus with a mass ratio of 2:1, and the rest parameters are the same as those of the method and the example 1.
Experimental example 1
The experimental examples explore the soothing and repairing effects of the ginsenoside extracts prepared in examples 1 to 3, and the steps are as follows: 70 female volunteers aged 20-50 years and having normal skin were selected and randomly and equally divided into 7 groups, namely, example 1-3 groups, comparative example 1-3 groups and control group. The skin of arms of volunteers is stimulated by using capsicum oil, left and right forearms of each group of volunteers are cleaned under the condition of room temperature and comfortable environment, a square area of 5cm multiplied by 5cm is respectively defined on the left and right forearms, 2mL of capsicum oil is smeared in the area, after the left and right forearms are stimulated for 20min, the capsicum oil is washed away, 2mL of ginsenoside extract of the corresponding group is smeared in the area coated with the capsicum oil respectively, and the area coated with the capsicum oil on the other side of the arm is reserved without smearing any product, so that the area is used as a blank control area. The control group was smeared with 2mL of clear water. The red and swollen condition of the forearms of the volunteers was observed at intervals and the time of complete analgesia of the forearms and the time of complete withdrawal of the redness were noted. The results are shown in Table 1.
TABLE 1 comparison of relief results for volunteers of each group
| Group of | Time/min required for pain relief | Time/min for red-removing |
| Example 1 | 27.9 | 53.4 |
| Example 2 | 30.8 | 55.5 |
| Example 3 | 29.2 | 53.4 |
| Comparative example 1 | 68.2 | 86.4 |
| Comparative example 2 | 73.8 | 95.1 |
| Comparative example 3 | 74.1 | 95.6 |
| Control | 85.6 | 115.4 |
As shown in Table 1, the ginsenoside extract prepared by the invention has the efficacy of relieving and repairing, can quickly relieve pain and redness of skin stimulated by capsicum oil after being used, and has the effect obviously superior to that of the comparative examples 1-3.
Experimental example 2
The experimental examples explore the anti-wrinkle whitening effect of the ginsenoside extracts prepared in examples 1 to 3.
1. Anti-wrinkle efficacy
70 female volunteers aged 20-50 years were selected and randomly and equally divided into 7 groups, examples 1-3, comparative examples 1-3 and control groups, respectively. The depth of wrinkles on the facial skin of each group of volunteers was observed using a VISIA-CR facial skin image analyzer, using the following experimental method: after each group of volunteers use clear water to clean the face, the corresponding group of products are taken to be coated on the face, the comparison group uses clear water to be coated on the face, the volunteers are massaged until the volunteers are completely absorbed, the number of times is 1/d, and the volunteers are continuously used for 28d. Wrinkle depth was observed with a VISIA-CR facial skin image analyzer before use at 1d and after use at 28d, and the wrinkle reduction rate after use of the product was counted for each group of volunteers. The results are shown in Table 2.
TABLE 2 comparison of anti-wrinkle results for groups of volunteers
| Group of | Wrinkle reduction rate/%after 28d use |
| Example 1 | 34.05 |
| Example 2 | 32.59 |
| Example 3 | 33.97 |
| Comparative example 1 | 20.11 |
| Comparative example 2 | 18.23 |
| Comparative example 3 | 21.74 |
| Control | 8.70 |
As shown in Table 2, the ginsenoside extract prepared by the invention has anti-wrinkle effect, can repair damaged collagen and collagen fibers after being used, and has the effect obviously superior to that of the comparative examples 1-3.
2. Whitening efficacy
70 female volunteers aged 20-30 years were selected and randomly and equally divided into 7 groups, examples 1-3, comparative examples 1-3 and control groups, respectively. The melanin content of the facial skin of each group of volunteers was determined by using the germany CK multi-probe skin test system MPA9, the experimental method being: after each group of volunteers use clear water to clean the face, the corresponding group of products are taken to be coated on the face, the comparison group uses clear water to be coated on the face, the volunteers are massaged until the volunteers are completely absorbed, the number of times is 1/d, and the volunteers are continuously used for 28d. After the 1 st, 7 th, 14 th, 21 st, 28 th products were used for 2h, their melanin content was measured with a probe at the same facial position. The results are shown in Table 3.
TABLE 3 comparison of the whitening results for each group of volunteers
As shown in Table 3, the ginsenoside extract prepared by the invention has the whitening effect, can effectively reduce melanin deposition in skin after being used, lightens skin color, and has the effect obviously superior to that of a comparative example group.
Experimental example 3
The experimental examples explore the anti-hair loss and hair growth effects of the ginsenoside extracts prepared in examples 1 to 3, and the steps are as follows: healthy SD rats were selected 70, and randomly and evenly divided into 7 groups, namely, examples 1 to 3 groups, comparative examples 1 to 3 groups and control groups, wherein after each group of rats is fed with basic feed for 7 days, the back hair of 3cm multiplied by 3cm on the back is removed, the corresponding ginsenoside extract is smeared on the exposed skin of the rats from the group 1 to 3 of the examples 1 to 3 of the depilation and the group 1 to 3 of the comparative examples, and physiological saline is smeared on the blank control group. The amount of the drug applied to each group of rats was 1 mL/time, 1 time/d, and 12d of drug was continuously applied. The growth of the back hair of the rat was recorded before each application from 5d, and the hair growth of the rat was classified into 5 grades, namely 0, 1, 2, 3 and 4, respectively, "0" means no hair growth, "1" means little hair growth, "2" means obvious hair growth, "3" means good hair growth, and "4" means normal hair growth. The results are shown in Table 4.
TABLE 4 comparison of rat dorsum hair growth conditions
As shown in Table 4, the ginsenoside extract prepared by the invention can grow villi on the 6 th to 7 th days after being smeared on the exposed skin of rats, is obviously earlier than that of the comparative example group and the control group, and can reach the normal level on the 11 th to 12 th days. In addition, the skin of the rats in the examples 1-3 does not have allergy phenomenon in the experimental process, and the activity and ingestion of the rats are not influenced, so that the ginsenoside extract prepared by the invention has no irritation and has the effects of preventing alopecia and promoting hair growth.
According to the embodiment, the invention provides the ginsenoside extract, the preparation method and the application thereof, and the ginseng is subjected to enzymolysis and fermentation, so that the extract contains various rare ginsenosides, and the product with effects of relieving and repairing, resisting wrinkles and whitening and preventing hair loss can be prepared.
The foregoing is merely a preferred embodiment of the present invention and it should be noted that modifications and adaptations to those skilled in the art may be made without departing from the principles of the present invention, which are intended to be comprehended within the scope of the present invention.
Claims (10)
1. A preparation method of a ginsenoside extract is characterized by comprising the following steps:
(1) Drying ginseng, pulverizing, mixing with glycosidase solution, and performing enzymolysis to obtain enzymolysis solution;
(2) Sterilizing the enzymolysis liquid, mixing with a composite microbial inoculum, and fermenting to obtain the ginsenoside extract.
2. The method according to claim 1, wherein the temperature of the drying in the step (1) is 40 to 50 ℃ and the time of the drying is 2 to 6 hours.
3. The method according to claim 1, wherein the ratio of ginseng to glycosidase solution in step (1) is 1g: 5-10 mL;
the glycosidase solution comprises one or more of beta-glucosidase, beta-galactosidase, alpha-arabinofuranosidase and beta-xylosidase;
the concentration of the enzyme in the glycosidase solution is independently 3-10U/mL.
4. The method according to claim 1, wherein the temperature of the enzymolysis in the step (1) is 40-50 ℃, and the time of the enzymolysis is 20-40 min.
5. The method according to claim 4, wherein the pH of the solution is adjusted to 4.5 to 5.5 before the enzymatic hydrolysis.
6. The preparation method according to claim 1, wherein the composite microbial inoculum in the step (2) comprises lactobacillus fermentum and lactobacillus rhamnosus, and the mass ratio of the lactobacillus fermentum to the lactobacillus rhamnosus is 1:1-3.
7. The method according to claim 1, wherein the inoculation amount of the composite microbial inoculum in the step (2) is 5-10%, the fermentation temperature is 35-40 ℃, and the fermentation time is 24-48 h.
8. The ginsenoside extract prepared by the preparation method of any one of claims 1 to 7.
9. The use of the ginsenoside extract of claim 8 for preparing products with effects of relieving, repairing, resisting wrinkle, whitening, preventing alopecia and promoting hair growth.
10. The use according to claim 9, wherein the type of product comprises a face cream, a serum cream, a face cream, a mask, a conditioner, a shampoo, a body cream.
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